A compound having the generalized structural formula or a pharmaceutically acceptable salt or prodrug thereof.

Full Text

FORM 2
THE PATENTS ACT 1970
[39 OF 1970]
&
THE PATENTS (AMENDMENT) RULES 2006
COMPLETE SPECIFICATION
[See Section 10; rule 13]
"SUBSTITUTED PYRIDINES AND PYRIDAZINES WITH ANGIOGENESIS INHIBITING ACTIVITY"
BAYER CORPORATION, 100 Bayer Road, Pittsburgh, Pennsylvania, 15205, U.S.A,
The following specification particularly describes the nature of the invention and the manner in which it is to be performed:-

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Substituted Pyridines and Pyridazines with Angiogenesis Inhibiting Activity
FIELD:
5 This application relates to small molecule heterocyclic pharmaceuticals, and more
particularly, to substituted pyridines and pyridazines having angiogenesis inhibiting activity.
BACKGROUND:
10 Vasculogenesis involves the de novo formation of blood vessels from endothelial
cell precursors or angioblasts. The first vascular structures in the embryo are formed by vasculogenesis. Angiogenesis involves the development of capillaries from existing blood vessels, and is the principle mechanism by which organs, such as the brain and the kidney are vascularized. While vasculogenesis is restricted to embryonic development, 15 angiogenesis can occur in the adult, for example during pregnancy, the female cycle, or wound healing.
One major regulator of angiogenesis and vasculogenesis in both embryonic development and some angiogenic-dependent diseases is vascular endothelial growth factor (VEGF; also called vascular permeability factor, VPF). VEGF represents a family 20 of mitogens isoforms resulting from alternative mRNA splicing and which exist in homodimeric forms. The VEGF KDR receptor is highly specific for vascular endothelial cells (for reviews, see: Farrara et al. Endocr. Rev. 1992, 13, 18; Neufield et al. FASEB J. 1999, 13, 9).
VEGF expression is induced by hypoxia (Shweiki et al. Nature 1992, 359, 843), as 25 well as by a variety of cytokines and growth factors, such as interleukin-1, interleukin-6, epidermal growth factor and transforming growth factor-a and -pTo date VEGF and the VEGF family members have been reported to bind to one or more of three transmembrane receptor tyrosine kinases (Mustonen et al. J. Cell Biol, 1995, 129, 895), VEGF receptor-1 (also known as fit-1 (fms-like tyrosine kinase-1)); 30 VEGFR-2 (also known as kinase insert domain containing receptor (KDR), the murine analogue of KDR being known as fetal liver kinase-1 (flk-1)); and VEGFR-3 (also known as flt-4). KDR and fit-1 have been shown to have different signal transduction properties (Waltenberger et al. J. Biol. Chem. 1994, 269, 26988); Park et al. Oncogene 1995, 10, 135). Thus, KDR undergoes strong ligand-dependent tyrosine phosphorylation in intact
1

1995, 72, 638), and age-related macular degeneration (AMD; see, Lopez et al. Invest. Opththalmol. Vis. Sci. 1996,37,855).
In rheumatoid arthritis (RA), the in-growth of vascular pannus may be mediated by
production of angiogenic factors. Levels of immunoreactive VEGF are high in the
5 synovial fluid of RA patients, while VEGF levels were low in the synovial fluid of
patients with other forms of arthritis of with degenerative joint disease (Koch et al. J.
Immunol. 1994, 152, 4149). The angiogenesis inhibitor AGM-170 has been shown to
prevent neovascularization of the joint in the rat collagen arthritis model (Peacock et al. J.
Exper. Med. 1992, 175, 1135).
10 Increased VEGF expression has also been shown in psoriatic skin, as well as
bullous disorders associated with subepidermal blister formation, such as bullous pemphigoid, erythema multiforme, and dermatitis herpetiformis (Brown et al. J. Invest. Dermatol. 1995, 104, 744).
Because inhibition of KDR signal transduction leads to inhibition of VEGF-15 mediated angiogenesis and permeabilization, KDR inhibitors will be useful in treatment of diseases characterized by abnormal angiogenesis and/or hyperpermeability processes., including the above listed diseases
Examples of phthalazines and other fused pyridazines that are similar in structure to those of the present application are disclosed in the following patents or patent 20 applications: WO 9835958 (Novartis), US 5,849,741, US 3,753,988, US 3,478,028 and JP 03106875. Other literature references to phthalazines are El-Feky, S.A., Bayoumy, B.E., and Abd El-Sami, Z.K., Egypt. J. Chem. (1991), Volume Date 1990, 33(2), 189-197; Duhault, J., Gonnard, P., and Fenard, S., Bull. Soc. Chirm Biol., (1967), 49 (2), 177-190; and Holava, H.M. and Jr, Partyka, R.A., J. Med. Chem., (1969), 12, 555-556. The 25 compounds of the present invention arc distinct from those described in each of the above references, and only the Novartis publication describes such compounds as inhibitors of angiogenesis.
As explained above, compounds which inhibit angiogenesis have applicability in treatment of a variety of medical conditions, and are therefore desirable. Such materials 30 are the subject of the present application.
SUMMARY:
In its broadest aspect, the present invention relates to the sum of three sets of chemical compounds, or pharmaceutically acceptable salts or prodrugs thereof, with each
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set overlapping the others in scope. The generalized structural formula for the compounds in each of the three sets of compounds is the same, but it should be noted that the definitions of the several groups comprising the' general structure in each set differ somewhat. Thus, the defined sets of chemical compounds differ from each other, but overlap in their scopes.
The first set of compounds have the generalized structural formula

■X— (CR4z
10

(G(I)
wherein
R1 and R2
together form a bridge containing two T? moieties and one T3 moiety, said bridge, taken together with the ring to which it is attached, forming a bicyclic of structure

T3 wherein each T2 independently represents N, CH, or CG4; T3 represents S, O, CR4G4', C(R4)2, or NR3; G4' represents any of the above-defined moieties G4 which are monovalent; and binding to ring J is achieved via terminal atoms T and T ;
b)
] 0 wherein each T2 independently represents N, CH, or CG4; G4' represents
any of the above-defined moieties G4 which are monovalent; with the proviso that a maximum of two bridge atoms T may be N ; and binding to
ring J is achieved via terminal atoms T ; and
c)

T4

T\; T5 J5- -T6 T5—
T6 ■L i T6 ■ or i
jV T^ Xs- -T3 >T5 T3
15
wherein each T4, T5, and T6 independently represents O, S, CR4G4', C(R4)2,
or NR3; G4' represents any of the above-defined moieties G4 which are
monovalent; and binding to ring J is achieved via terminal atoms T4 or T5;
with the provisos that:
20 i) when one T4 is O, S, or NR3, the other T4 is CR4G4' or C(R4)2 ;
' ii) a bridge comprising Ts and T atoms may contain a maximum of
two heteroatoms O, S, or N; and
iii) in a bridge comprising T5 and T atoms, when one T group and
one T group are 0 atoms, or two T groups are O atoms, said O
25 atoms are separated by at least one carbon atom.

4 >
When G4 is an alkyl group located on ring J adjacent to the linkage -(CR'
and X is NR3 wherein RJ is an alkyl substituent, then G4 and the alkyl substituent RJ on X may be joined to form abridge of structure -(CH2)P'- wherein p' is 2, 3, or 4, with
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the proviso that the sum of p and p. is 2, 3, or 4, resulting in formation of a nitrogen-containing ring of 5, 6, or 7 members.
Additional provisos are that: 1) in G1, G2, G3, and G4, when two groups R3 or
5 R6 are each alkyl and located on the same N atom they may be linked by a bond, an O,
an S, or NR to form a N-containing heterocycle of 5 - 7 ring atoms; and 2) when an
aryl, heteroaryl, or heterocyclyl ring is optionally substituted, that ring may bear up to
5 substituents which are independently selected from the group consisting of amino,
mono-loweralkyl-substituted amino, di-loweralkyl-substituted amino, lower
10 alkanoylamino, halogeno, lower alkyl, halogenated lower alkyl, hydroxy, lower
alkoxy, lower alkylthio, halogenated lower alkoxy, halogenated lower alkylthio, lower
alkanoyloxy, -C02R3, -CHO, -CH2OR3, -OC02R3, -CON(R6)2, ~OCON(R6)2,
-NR3CON(R6)2 , nitro, amidino, giianidino, mercapto, sulfo, and cyano; and 3) when
any alkyl group is attached to O, S, or N, and bears a hydroxyl substituent, then said
15 hydroxyl substituent is separated by at least two carbon atoms from the O, S, or N to
which the alkyl group is attached.
The second set of compounds have the generalized structural formula

D^E

(G3)q

(I)

20 wherein
R1 and R2:
i) independently represent H or lower alkyl; ii) together form a bridge of structure
G1)(
25 wherein binding is achieved via the terminal carbon atoms;
iii) together form a bridge of structure
G1),

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wherein binding is achieved via the terminal carbon atoms; IV) together form a bridge of structure T1
r
V
=r
wherein one or two ring members T1 are N and the others are CH or CG1, and binding is achieved via the terminal atoms; or v) together form a bridge containing two T2 moieties and one T3 moiety, said bridge, taken together with the ring to which it is attached, forming a bicyclic of structure

b)
^T2
wherein each T independently represents N, CH, or CG ; G4' represents any of the above-defined moieties G which are monovalent; with the proviso that a maximum of two bridge atoms T may be N ; and binding to ring J is achieved via terminal atoms T2; and c)

T4 \« T5^T5 Ts- -T6 T5—Tv6T6
| T6 L I T6 , or !
T4 T*/ ^ T5- ^T6 J6T5—T6
wherein each T4, T5, and T6 independently represents O, S, CR4G4, C(R4)2,
10 or NR ; G4' represents any of the above-identified moieties G4 which are
monovalent; and binding to ring J is achieved via terminal atoms F or T5 ;■ with the provisos that:
i) when one T4 is O, S, orNR3, the other T4 is CR4G4'or C(R4)2 ;
ii) a bridge comprising T and T atoms may contain a maximum of
15 two heteroatoms O, S, or N; and
iii) in a bridge comprising T5 and T atoms, when one T5 group and one T group are O atoms, or two T groups are O atoms, said O atoms are separated by at least one carbon atom.
20 . When G* is an alkyl group located on ring J adjacent to the linkage -(CR42)P- ,
and X is NR3 wherein R3 is an alkyl substituent, then G4 and the alkyl substituent R3 on X may be joined to form a bridge of structure -(CH2)P' - wherein p' is 2, 3, or 4, with the proviso that the sum of p and p' is 2, 3, or 4, resulting in formation of a nitrogen-containing ring of 5, 6, or 7 members.
25
Additional provisos are that: 1) in G1, G2, G3, and G4, when two groups R3 or R6 are each alkyl and located on the same N atom they may be linked by a bond, an O, an S, or NR3 to form a N-containing heterocycle of 5 - 7 ring atoms; and 2) when an aryl, heteroaryl, or heterocyclyl ring is optionally substituted, that ring may bear up to 5 substituents which are independently selected from the group consisting of amino,
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mono-loweralkyl-substituted amino,. di-loweralkyl-substituted amino, lower alkanoylamino, halogeno, lower alkyl, halogenated lower alkyl, hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy, halogenated lower alkylthio,- lower alkanoyloxy, -C02R3, -CHO, -CH2OR3, -OC02R3, -CON(R6)2, -OCON(R6)2) -NR3CON(R6)2, nitro, amidino, guanidino, mercapto, sulfo, and cyano; and 3) when any alkyl group is attached to O, S, or N, and bears a hydroxyl substituent, then said hydroxyl substituent is separated by at least two carbon atoms from the O, S, or N to which the alkyl group is attached.
10
The third set of compounds have the generalized structural formula

X— (CR42V—(- J
,N-^. P
z' VR1 (G
wherein
R1 and R2:
15 i) independently represent H or lower alkyl;
ii) together form a bridge of structure
^ ) m
wherein binding is achieved via the terminal carbon atoms; lii) together form a bridge of structure
20 G >™
wherein binding is achieved via the terminal carbon atoms;
iv) together form a bridge of structure
'V
wherein one or two ring members T1 are N and the others are CH or CG1, and
25 binding is achieved via the terminal atoms; or
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wherein each T2 independently represents N, CH, or CG ; G4' represents any of the above-defined moieties G4 which are monovalent; with the proviso that a maximum of two bridge atoms T may be N ; and binding to
ring J is achieved via terminal atoms T ; and
c)

T4 ^e T5- -T6 T5___T6T6
| T6 L J T6 , or I
T4 T5/ T^1* T5- / ^T6 >T5—Ts
wherein each T4, T5, and T6 independently represents 0, S, CR4G4', C(R4)2,
or NR3; G4' represents any of the above-defined moieties G4 which are
.monovalent; and binding to ring J is achieved via terminal atoms rorT5;
10 with the provisos that:
■ i) when one T4 is O, S, or NR3, the other T4 is CR4G4' or C(R4)2; ii) a bridge comprising Ts and T atoms may contain a maximum of
two heteroatoms O, S, or N; and
iii) in a bridge comprising T and T atoms, when one T group and
15 one T6 group are O atoms, or two T6 groups are O atoms, said O
atoms are separated by at least one carbon atom;
When G4 is an alkyl group located on ring J adjacent to the linkage -(CR42)P- ,
and X is NRJ wherein RJ is an alkyl substituent, then G* and the alkyl substiruent RJ
20 on Xmay be joined to form a bridge of structure -(CH2)P'- wherein p' is 2, 3, or 4, with
the proviso that the sum of p and p' is 2, 3, or 4, resulting in formation of a nitrogen-containing ring of 5, 6, or 7 members.
Additional provisos are that: 1) in G1, G2, G3, and G4, when two groups R3 or R6 are
25 each alkyl and located on the same N atom they may be linked by a bond, an O, an S,
or NR3 to form a N-containing heterocycle of 5 - 7 ring atoms; and 2) when an aryl,
heteroaryl, or heterocyclyl ring is optionally substituted, that ring may bear up to 5
substituents which are independently selected from the group consisting of amino,
mono-loweralkyl-substituted amino, di-loweralkyl-substituted amino, lower
30 alkanoyiamino, halogeno, lower alkyl, halogenated lower alkyl, hydroxy, lower
alkoxy, lower alkylthio, halogenated lower alkoxy, halogenated lower alkylthio, lower alkanoyloxy, -C02R3, -CHO, -CH2OR3, -OCQ2R3, -CON(R6)2, -OCON(R6)2,
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-NR3CON(R6)2, nitro, amidino, guanidino, mercapto, sulfo, and cyano; and 3) when any alkyl group is attached to O, S, or N, and bears a hydroxyl substituent, then said hydroxyl substituent is separated by at least two carbon atoms from the 0, S, or N to which the alkyl group is attached. 5
Pharmaceutically acceptable salts of these compounds as well as commonly used
prodrugs of these compounds such as O-acyl derivatives of invention compounds which
contain hydroxy groups are also within the scope of the invention.
10 The invention also relates to pharmaceutical compositions comprising one or more
of the compounds of the invention, or their salts or prodrugs, in a pharmaceutically
acceptable carrier.
The invention also relates to a method for using these materials to treat a mammal
having a condition characterized by abnormal angiogenesis or hyperpermiability 15 processes, comprising administering to the mammal an amount of a compound of the
invention, or a salt or prodrug thereof, which is effective to treat the condition.
DETAILED DESCRIPTION:
20 Definitions:
The prefix "lower" denotes a radical having up to and including a maximum of 7 atoms, especially up to and including a maximum of 5 carbon atoms, the radicals in question being either linear or branched with single or multiple branching.
"Alkyl" means a hydrocarbon radical having up to a maximum of 12 carbon 25 atoms, which may be linear or branched with single or multiple branching. Alkyl is especially lower alkyl.
Where the plural form is used for compounds, salts, and the like, this is taken to mean also a single compound, salt, or the like.
Any asymmetric carbon atoms may be present in the (R)-, (S)- or
30 (Reconfiguration, preferably in the (R)- or (S)-configuration. Substituents at a double
bond or a ring may be present in cis- (= Z-) or trans (= E-) form. The compounds may
thus be present as mixtures of isomers or as pure isomers, preferably as enantiomer-pure
diastereomers and having pure cis- or trans- double bonds.
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Lower alkylene Y may be branched or linear but is preferably linear, especially methylene (-CH2), ethylene (-CH2-CH2), trimethylene (-CH2-CH2-CH2) or tetramethylene (-CH2CH2CH2CH2). When Y is lower alkylene, it is most preferably methylene.
"Aryl" means an aromatic radical having 6 to 14 carbon atoms, such as phenyl, 5 naphthyl, fluorenyl or phenanthrenyl.
"Halogen" means fluorine, chlorine, bromine, or iodine but is especially fluorine, chlorine, or bromine.
"Pyridyl" means 1-, 2-, or 3-pyridyl but is especially 2- or 3-pyridyl. "Cycloalkyl" is a saturated carbocycle that contains between 3 and 12 carbons but 10 preferably 3 to 8 carbons.
"Cycloalkenyl" means a non-reactive and non-aromatic unsaturated carbocycle that contains between 3 and 12 carbons but preferably 3 to 8 carbons and up to three double bonds. It is well known to those skilled in the art that cycloalkenyl'groups that differ from aromatics by lacking only one double bond such as cyclohaxadiene are not 15 sufficiently non-reactive to be reasonable drug substances and therefor their use as substituents is not within the scope of this invention.
Cycloalkyl and cycloalkenyl groups may contain branch points such that they are
substituted by alkyl or alkenyl groups. Examples of such branched cyclic groups are 3,4-
dimethylcyclopentyl, 4-allylcyclohexyI or 3-ethylcyclopent-3-enyl.
20 Salts are especially the pharmaceutically acceptable salts of compounds of formula
I such as, for example, acid addition salts, preferably with, organic or inorganic acids, from compounds of formula I with a basic nitrogen atom. Suitable inorganic acids are, for example, halogen acids such as hydrochloric acid, sulfuric acid, or phosphoric acid. Suitable organic acids are, for example, carboxylic, phosphonic, sulfonic, or sulfamic 25 acids, for example acetic acid, propionic acid, octanoic acid, decanoic acid, dodecanoic acid, glycolic acid, lactic acid, -hydroxybutyric acid, gluconic acid, glucosemonocarboxylic acid, fumaric acid, succinic acid, adipic acid, pimelic acid, suberic acid, azeiaic acid, malic acid, tartaric acid, citric acid, glucaric acid, galactaric acid, amino acids, such as glutamic acid, aspartic . acid, N-methylglycine, 30 acetytaminoacetic acid, N-acetylasparagine orN-acetylcysteine, pyruvic acid, acetoacetic acid, phosphoserine, 2- or 3-glycerophosphoric acid.
In the definition of Y, the diradical "-(5 member heteroaryl)-" denotes a 5-membered aromatic heterocycle containing 1-3 heteroatoms selected.from 0, S, and N, the number of N atoms being 0-3 and the number of O and S atoms each being 0-1 and
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connected to the sulfur from a carbon and to -(CR 2)s- through a C or N atom. Examples
of such diradicals include .

v \N H
N '/ V
O
n,
N H

N-N
7 V
N"
H
// W
V
N H

N \\ 7 V
o
N"w
7 \
In the definitions of G1, G2, G3, and C the statement is made that when two groups R3 or R6 are found on a single N, they can be combined into a heterocycle of 5-7 atoms. Examples of such heterocycles, including the N to which they are attached, are:
CH3

-N

-N

O

-N

—N

O
t

-N

N-RJ

2n5
CM.
2n5
C,H.

,RJ

CH3

—N

O

"Heterocyclyl" or "heterocycle" means a five- to seven-membered heterocyclic system with 1-3 heteroatoms selected from the group nitrogen, oxygen, and sulfur, which 10 may be unsaturated or wholly or partly saturated, and is unsubstituted or substituted especially by lower alkyl, such as methyl, ethyl, 1-propyl, 2-propyl, or tert-butyl.
When an aryl, heteroaryl, or heterocyclyl ring is said to be optionally substituted,
that ring may bear up to 5 substituents which are independently selected from the group
consisting of amino, mono- or di-loweralkyl-substituted amino, lower alkanoylamino,
15 halogeno, lower alkyl, halogenated lower alkyl such as trifluoromethyl, hydroxy, lower
alkoxy, lower alkylthio, halogenated lower alkoxy such as trifluoromethoxy, halogenated
lower alkylthio such as trifluoromethylthio, lower alkanoyloxy, -C02R3, -CHO, -CH2OR3,
-OC02R3, -CON(R6)2, -OCO N(R6)2) -NR3CON(R6)2, nitro, amidino, guanidmo,
mercapto, sulfo, and cyano.
20 In the ring attached to Y, the ring members A, B, D, E, and L may be N or CH, it
being understood that the optional substituents G3 are necessarily attached to carbon and
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not nitrogen, and that when a given carbon bears a substituent group G , that G3 group is in place of the H atom the carbon would bear in the absence of the G3 group.

"Heteroaryl" means a monocyclic or fused bicyclic aromatic system with between 5 and 10 atoms in total of which 1-4 are heteroatoms selected from the group comprising nitrogen, oxygen, and sulfur and with the remainder being carbon. Heteroaryl is preferably a monocyclic system with 5 or 6 atoms in total, of which 1-3 are heteroatoms.
10 "Alkenyl" means an unsaturated radical having up to a maximum of 12 carbon
atoms and may be linear or branched with single or multiple branching and containing up
to 3 double bonds. Alkenyl is especially lower alkenyl with up to 2 double bonds.
"Alkanoyl" means alkylcarbonyl, and is especially lower alkylcarbonyl.
Halogenated lower alkyl, halogenated lower alkoxy and halogenated lower
15 alkylthio are substituents in which the alkyl moieties are substituted either partially or in
full with halogens, preferably with chlorine and/or fluorine and most preferably with
fluorine. Examples of such substituents are trifluoromethyl, trifluoromethoxy,
trifluoromethylthio, 1,1,2,2-tetrafluoroethoxy, dichlororaethyl, flu orom ethyl and
difluoromethyl.
20 When a substituent is named as a string of fragments such as "phenyl-lower
alkoxycarbonyl-substituted alkylamino," it is understood that the point of attachment is to
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the final moiety of that string (in this case amino) and that the other fragments of that string are connected to each other in sequence as they are listed in the string. Thus an example of "phenyl-lower alkoxycarbonyl-substituted alkylamino" is:
o ""'
O j
point of attachment from nitrogen
amino
carbonyl
f
phenyl
lower alkoxy alkyl
When a substituent is named as a string of fragments with a bond at the start (typically written as a dash) such as "-S(0)p(optionally substituted heteroarylalkyl)", it is understood that the point of attachment is to the first atom of that string (in this case S or sulfur) and that the other fragments of that string are connected to each other in sequence as they are listed in the string. Thus an example of "-S(0)p(optionally substituted heteroarylalkyl)" is:

F3C~
optional ^ substituent

heteroaryl

>°.
S(0)p

point of attachment from sulfur

alkyl
It is to be understood that the left-most moiety of each of the variants of the linker Y is connected to the ring containing A, B, D, E, and L and that the right-most moiety of the linker is connected to the pyridazine fragment of the generalized formulae. Thus, examples of the use of the linker "-CH2-0-" or of the linker "-0-CH2-" are represented in the following invention compounds:

H9N

In generalized structural formula (I), the preferred and most preferred groups are as follows.

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R1 and R2 preferably: ' i) together form a bridge of structure

G1)m
wherein binding is achieved via the terminal carbon atoms; or ii) together form a bridge of structure
V
T1=T1 wherein one of the ring members T is N and the others are CH, and binding is achieved via the terminal atoms; or iii) together form a bridge containing two T2 moieties and one T3 moiety, said bridge, taken together with the ring to which it is attached, forming a bicyclic of structure

T\, T5^ -J6
T6 ' r or T6
T5/ T6 T5- /
20
wherein each T5, and T5 independently represents O, S, or CH2 ; and
binding to ring J is achieved via terminal atoms T ; with the provisos that:
i) a bridge comprising T5 and T6 atoms may contain a maximum of
two heteroatoms O, S, or N; and
25 ii) in a bridge comprising T5 and T6 atoms, when one T5 group and
one T6 group are O atoms, or two T groups are Q atoms, said O
atoms are separated by at least one carbon atom.
Alkyl groups which constitute all or part of a G4 moiety are preferably lower alkyl.
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When G4 is an alkyl group located on ring J adjacent to the linkage -(CR '2)p-, and X is NR3 wherein R3 is an alkyl.substituent, then G4 and the alkyl substituent R3 on X may be joined to form a bridge of structure -(CH2)P'- wherein p' is preferably 2 or 3, with the proviso that the sum of p and p' is 2 or 3, resulting in formation of a nitrogen-containing 5 ring of 5 or 6 members. Most preferably, the sum of p and p' is 2, resulting in formation of a 5-membered ring.
Most preferably, in G1, G2, G3, and G4, when two groups R6 are each alkyl and
located on the same N atom they may be linked by a bond, an O, an S, or NR3 to form a
N-containing heterocycle of 5 — 6 ring atoms.
10 Preferably, when an aryl, heteroaryl, or heterocyclyl ring is optionally substituted,
that ring may bear up to 2 substituents which are independently selected from the group consisting of amino, mono-loweralkyl-substituted amino, di-Ioweralkyl-substituted amino, lower alkanoylamino, halogeno, lower alkyl, halogenated lower alkyl, hydroxy, lower alkoxy, lower alkylthio, halogenated lower alkoxy, halogenated lower alkylthio, -CH2OR3, 15 nitro, and cyano.
The method of the invention is intended to be employed for treatment of VEGF-mediated conditions in both humans and other mammals.
The compounds may be administered orally, dermally, parenterally, by injection,
by inhalation or spray, or sublingually, rectally or vaginally in dosage unit formulations.
20 The term 'administered by injection' includes intravenous, intraarticular, intramuscular,
subcutaneous and parenteral injections, as well as use of infusion techniques. Dermal
administration may include topical application or transdermal administration. One or
more compounds may be present in association with one or more non-toxic
pharmaceutically acceptable carriers and if desired, other active ingredients.
25 Compositions intended for oral use may be prepared according to any suitable
method known to the art for the manufacture of pharmaceutical compositions. Such
compositions may contain one or more agents selected from the group consisting of
diluents, sweetening agents, flavoring agents, coloring agents and preserving agents in
order to provide palatable preparations.
30 Tablets contain the active ingredient in admixture with non-toxic pharmaceutically
acceptable excipients which are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; and binding agents, for example magnesium stearate,
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stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to' delay disintegration and adsorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material" such as glyceryl monostearate or glyceryl distearate may be employed. These compounds may 5 also be prepared in solid, rapidly released form.
Formulations for oral use may also be presented as hard gelatin capsules wherein
the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate,
calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is
mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil.
10 Aqueous suspensions containing the active materials in admixture with excipients
suitable for the manufacture of aqueous suspensions may also be used. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropyl-methylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanfh and gum acacia; dispersing or wetting agents may be a naturally-occurring phosphatide, 15 for example, lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with 20 paitial esters derived from fatty acids and hexitol anhydrides, for example polyethylene
» *v»-l— .in .^ .T-. ^-. —. —. r-\ 1 r* r-t +-*> " V* o " m t o"~»" I r* nun »^ t~\+*
&\JL ULLUll lllulluULtaLC. X 1J.C aljutuuo ouiop^iioluiio tLicij' aiiu vwiiium VJIAV. \JL aiui^
preservatives, for example ethyl, or «-propyl, y?-hydroxybenzoate, one or more coloring
agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose
or saccharin.
25 Dispersiblc powders and granules suitable for preparation of an aqueous
suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example, sweetening, flavoring and coloring 30 agents, may also be present.
The compounds may also be in the form of non-aqueous liquid formulations, e.g., oily suspensions which may be formulated by suspending the active ingredients in a vegetable oil, for example arachis oil, olive oil, sesame oil or peanut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example
27

WO 01/23375 PCT/US00/26500
beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide palatable oral preparations. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.
Pharmaceutical compositions of the invention may also be in the form of oil-in-5 water emulsions. The oil phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures of these. Suitable emulsifying agents may be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, 10 and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents.
Syrups and elixirs may be formulated with sweetening, agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a 15 demulcent, a preservative and flavoring and coloring agents.
The compounds may also be administered in the form of suppositories for rectal or
vaginal administration of the drug. These compositions can be prepared by mixing the
drug with a suitable non-irritating excipient which is solid at ordinary temperatures but
liquid at the rectal or vaginal temperature and will therefore melt in the rectum or vagina
20 to release the drug. Such materials include cocoa butter and polyethylene glycols.
Compounds of the invention may also be administered transdermally using methods known to those skilled in the art (see, for example: Chien; "Transdermal Controlled Systemic Medications"; Marcel Dekker, Inc.; 1987. Lipp et al. WO 94/04157 3Mar94). For example, a solution or suspension of a compound of Formula I in a suitable 25 volatile solvent optionally containing penetration enhancing agents can be combined with additional additives known to those skilled in the art, such as matrix materials and bacteriocides. After sterilization, the resulting mixture can be formulated following known procedures into dosage forms. In addition, on treatment with emulsifying agents and water, a solution or suspension of a compound of Formula I may be formulated into a 30 lotion or salve.
Suitable solvents for processing transdermal delivery systems are known to those skilled in the art, and include lower alcohols such as ethanol or isopropyl alcohol, lower ketones such as acetone, lower carboxylic acid esters such as ethyl acetate, polar ethers such as tetrahydrofuran, lower hydrocarbons such as hexane, cyclohexane or benzene, or
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halogenated hydrocarbons such as dichloromethane, chloroform, trichlorotrifluoroethane, or trichlorofluoroethane. Suitable solvents may also include mixtures one or more materials selected from lower alcohols, lower ketones , lower carboxylic acid'esters, polar ethers, lower hydrocarbons, halogenated hydrocarbons.
Suitable penetration enhancing materials for transdermal delivery systems are known to those skilled in the art, and include, for example, monohydroxy or polyhydroxy alcohols such as ethanol, propylene glycol or benzyl alcohol, saturated or unsaturated Cg-Ci8 fatty alcohols such as lauryl alcohol or cetyl alcohol, saturated or unsaturated Cg-Cis fatty acids such as stearic acid, saturated or unsaturated fatty esters with up to 24 carbons such as methyl, ethyl, propyl, isopropyl, rc-butyl, sec-butyl isobutyl tert-butyl or monoglycerin esters of acetic acid, capronic acid, lauric acid, myristinic acid, stearic acid, or palmitic acid, or diesters of saturated or unsaturated dicarboxylic acids with a total of up to 24 carbons such as diisopropyl adipate, diisobutyl adipate, diisopropyl sebacate, diisopropyl maleate, or diisopropyl fumarate. Additional penetration enhancing materials include phosphatidyl derivatives such as lecithin or cephalin, terpenes, amides, ketones, ureas and their derivatives, and ethers such as dimethyl isosorbid and diethyleneglycol monoethyl ether. Suitable penetration enhancing formulations may also include mixtures one or more materials selected from monohydroxy or polyhydroxy alcohols, saturated or unsaturated Cg-Cis fatty alcohols, saturated or unsaturated Cg-Cis fatty acids, saturated or unsaturated fatty esters with up to 24 carbons, diesters of saturated or unsaturated dicarboxylic acids with a total of up to 24 carbons, phosphatidyl derivatives, terpenes, amides, ketones, ureas and their derivatives, and ethers.
Suitable binding materials for transdermal delivery systems are known to those skilled in the art and include polyacrylates, silicones, polyurethanes, block polymers, styrene-butadiene coploymers, and natural and synthetic rubbers. Cellulose ethers, derivatized polyethylenes, and silicates may also be used as matrix ■ components. Additional additives, such as viscous resins or oils may be added to increase the viscosity of the matnx.
For all regimens of use disclosed herein for compounds of Formula I, the daily oral dosage regimen will preferably be from 0.01 to 200 mg/Kg of total body weight. The daily dosage for administration by injection, including intravenous, intramuscular, subcutaneous and parenteral injections, and use of infusion techniques will preferably be from 0.01 to 2.00 mg/Kg of total body weight. The daily rectal dosage regimen will preferably be from 0.01 to 200 mg/Kg of total body weight. The daily vaginal dosage regimen will
29

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preferably be from 0.01 to 200 mg/Kg of total body weight. The daily topical dosage regimen will preferably be from 0.1 to 200 mg administered between one to four times daily. The transdermal concentration will preferably be that required to maintain a daily dose of from 0.01 to 200 mg/Kg. The daily inhalation dosage regimen will preferably be 5 from 0.01 to 10 mg/Kg of total body weight.
It will be appreciated by those skilled in the art that the particular method of administration will depend on a variety of factors, all of which are considered routinely when administering therapeutics. It will also be understood, however, that the specific dose level for any given patient will depend upon a variety of factors, including, but not 10 limited to the activity of the specific compound employed, the age of the patient, the body weight of the patient, the general health of the patient, the gender of the patient, the diet of the patient, time of administration, route of administration, rate of excretion, drug combinations, and the severity of the condition undergoing therapy. It will be further appreciated by one skilled in the art that the optimal course of treatment, i.e., the mode of 15 treatment and the daily number of doses of a compound of Formula I or a pharmaceutical^ acceptable salt thereof given for a defined number of days, can be ascertained by those skilled in the art using conventional treatment tests.
GENERAL PREPARATIVE METHODS
20
The compounds of the invention may be prepared by use of known chemical
reactions and procedures. Nevertheless, the following general preparative methods are
presented to aid the reader in synthesizing the KDR inhibitors, with more detailed
particular, examples being presented below in the experimental section describing the
25 working examples.
All variable groups of these methods are as described in the generic description if
they are not specifically defined below. When a variable group or substiruent with a given
symbol (i.e. R3, R4, R , G1, G2, G3, or G4) is used more than once in a given structure, it is
to be understood that each of these groups or substituents may be independently varied
30 within the range of definitions for that symbol. As defined above, the compounds of the
invention contain ring units each of which may independently bear between 0 and 5
substituents G1, G\ or G , which are not defined as H. By contrast, it is to be noted that in
the general method schemes below, the G , G3, or G4 substituents are used as if their
definition includes H, to show where such G1, GJ, or G4 substituents may exist in the
30

WO 01/23375 PCT7USO0/265O0
' , -, .
structures, and for ease in drawing. No change in the definition of G , G , or G is
intended by this non-standard usage, however. Thus, only for purposes of the general
method schemes below, G , G , or G may be H in addition to the moieties set forth in the
definitions of G1, G3, or G . The ultimate compounds contain 0 to 5 non-hydrogen groups
G', G3, or G4.
Within these general methods the variable M is equivalent to the moiety
-(CR42)p-(7(W)q
—' in which each variable group or substituent is allowed to
independently vary within the limits defined earlier for that symbol.
10 Within these general methods the variable Q is equivalent to the moiety
A-B
q in which L is N and each other variable group or substituent is
allowed to independently vary within the limits defined earlier for that symbol.
Within these general methods the variable Q2 is equivalent to the moiety
A-B L7/ (V-b=E>G3}
Q in which each variable group or substituent is allowed to
15 independently vary within the limits defined earlier for that symbol.
It is recognized that compounds of the invention with each claimed optional
functional group cannot be prepared with each of the below-listed methods. Within the
scope of each method optional substituents are used which are stable to the reaction
conditions, or the functional groups which may participate in the reactions are present in
20 protected form where necessary, and the removal of such protective groups is completed
at appropriate stages by methods well known to those skilled in the art.
General Method A - The compounds of formula I-A in which X, M, and Q~ are
defined as above, Y is -CH2-0-, -CH2-S-, -CH2-NH-, -0-, -S-, or -NH-, and R1 and R2 25 together with the carbons to which they are attached form a fused 5-membered- ring aromatic heterocycle, hal is halogen (CI, Br, F, or I but preferably CI, Br or F) are conveniently prepared according to a reaction sequence as shown in Method A. Thus, a heterocycle of formula II in which R is lower alkyl can be made by one skilled in the art according to the corresponding published procedures in the reference table. In the cases 30 of thiophene-2,3-dicarboxylic acid (table entry 1) and pyrazole-3,4-dicarboxy!ic acid
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(table entry 10), the carboxylic acids are converted to methyl or ethyl esters by treatment with the corresponding alcohol and catalytic mineral acid (typically sulfuric acid) at reflux. The diester of formula II is treated with hydrazine hydrate to furnish intermediate III (for specific reaction conditions see Robba, M.; Le Guen, Y. Bull. Soc. Chem. Fr.. 5 1970 12 4317). Compound III is treated with a halogenating agent such as phosphorous oxychloride, phosphorous oxybromide, phosphorous pentabromide, or phosphorous pentachloride to yield dihalo intermediate IV. The dichloro or dibromo intermediates can be converted to the difluoro intermediate (when desired) by reaction with hydrogen fluoride. By using iodo reagents such as potassium iodide or tetrabutylammonium iodide 10 in subsequent steps, the iodo intermediate is formed in the reaction mixtures without being isolated as a pure substance. Dihalo intermediate IV is treated with a nucleophile of formula V in refluxing alcohol or other suitable solvent such as tetrahydrofuran (THF), dimethoxyethane (DME), dimethylformamide (DMF), dimethylsulfoxide (DMSO), or the like to furnish the intermediate of formula VI. Such condensations can also be done in a 15 melt free of solvent and can be catalyzed by acids such as HCl or bases such as triethylamine or l,8-diazobicyclo[5.4.0]undec-7-ene (DBU). The compound of formula VI is reacted with compounds of formula VII in a suitable aprotic solvent such as DMSO, DMF or solvent free often with a basic catalyst such as DBU or CsCCv, or a crown ether such as 18-crown-6 at temperatures usually between room temperature and reflux to 20 furnish invention compound of formula I-A. It is understood that the nature of the starting materials will dictate the choice of suitable solvents, catalyst (if used) and temperature by one skilled in the art. Intermediates of formula V and VII are often commercial or are conveniently prepared by methods well known to those skilled in the art. For example see Martin, I., et al. Acta. Chem. Scand.. 1995 49 230 for the preparation of VII in which Y is 25 -CH2-O- and Q2 is 4-pyridyl substituted by a 2-aminocarbonyl group (2-CONH2).
32

Et02C H Nicolaus, Mangoni. Gazz. Chim. Ital. 1956 55 757.
Et02ChC0!E' Sice, J. J. Org. Chem.. 1954 19 70.
KNXN^C02Et Tanaka, Y. Tetrahedron. 1973 29 3271.
C02CH3NCXj_l C02CH3 Diacid: Tyupalo, N.; Semenyuk, T.; Kolbasina, O. ^z«x /. PAys. Chem. 1992
General Method B - The compounds of formula I-B in which M, X, and Q2 are as defined above and Y is -CH2-0-, -CH2-S-, -CH2-NH-, -O-, -S-, or -NH-. are conveniently prepared as shown in Method B. According to a procedure described in the literature (Tomisawa and Wang, Chem. Pharm. Bull, 21, 1973, 2607, 2612), isocarbostyril VIII is reacted with PBrs in a melt to form 1,4-dibromoisoquinoline IX. Intermediate IX is treated with, a nucleophile of formula V in refluxing alcohol to furnish intermediate of formula X. Such condensations can also be done in a melt free of solvent and can be catalyzed by acids such as HC1 or bases such as triethylamine or 1,8-diazobicyclo[5.4.0]undec-7-ene (DBU). The compound of formula X is reacted with compounds of formula VII in a suitable aprotic solvent such as DMSO, DMF or solvent free often with a basic catalyst such as DBU or CSCO4 at elevated temperatures to furnish invention compound of formula I-B. This method is most useful when Y is -CH2-S- or -S-.
34

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Method B
General Method C - The compounds of formula I-C in which M, X, R.1, R2, m and Q2 are defined as above are conveniently prepared according by a reaction sequence 5 as shown in method C. In this method m is preferably 0 and R and R2 together with the carbons to which they are attached form a fused benzene or fused 5-member ring aromatic heterocycle. Starting material XI is either commercial or is prepared by one skilled in the art as shown in the reference table below. Starting material XI is reacted with urea or ammonia, usually at elevated temperature and pressure (in the case of ammonia), to form 10 imide XII. The imide is reacted with an aldehyde XIII in acetic acid "and piperidine at reflux to yield intermediate XIV. Reaction of XIV with sodium borohydride in methanol or other suitable solvents according to the general procedure described by I.W. Elliott and Y. Takekoshi (J. Heterocyclic Chem. 1976 13, 597) yields intermediate XV. Treatment of XV with a suitable halogenatmg agent such as POCl3, POBr3, PCI5, PBr5 or thionyl 15 chloride yields halo intermediate XVI which is reacted with nucleophile of formula V in refluxing alcohol to furnish invention compound of formula I-C. Such condensations can also be done in a melt free of solvent and can be catalyzed by acids such as HC1 or bases such as triethylamine or l,8-diazobicyclo[5.4.0]undec-7-ene (DBU). Alternatively, reagent V can be condensed with intermediate XV be heating the two components with 20 P2O5 in a melt to yield invention compound of structure I-C. This last method is especially effective when X is an amine linker.
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XI

Urea
or NH,

XII

NH
O

o
(CH2)iSH Q XIII

(CH2)m XIV Q2

NaBH4

V
REFERENCE TABLE FOR PREPARATION OF STARTING MATERIALS

NH
O

"O

Commercial

Commercial

D.E. Ames and O. Ribeiro, J.Chem.Soc, Per/tin Trans. 1 1975, 1390.

J.R. Carson and S. Wong, J. Med. Chem. 1973, 16, 172.

O

"OH .OH

K. Yasuyuki, et al., J. Org. Chem. 1986, 51, 4150.

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WO 01/23375

PCT/USOO/26500

NH
0

O

Schneller, et al., J. Med. Chem. 1978, 21, 990.

R.K. Robins et al, J. Org. Chem. 1963, 28, 3041.

P. Gupta, et al., J. Heterocycl. Chem. 1986, 23, 59.

O

R. B. Meyer, et al, J. Heterocycl. Chem. 1980 17, 159.

N' H

OMe
OMe O

10

General Method D - The compounds of formula I-D-l in which R1, R , Rs, M, X,
Y, G3 and Z are defined as above and q is 0 or 1 are conveniently prepared via a reaction sequence as shown in Method D. Thus, pyridine substituted pyridazines or pyridines (I-D-1) are functionalized into substituted 2-aminocarbonyl pyridines of formula (I-D-2) by the use of formamides (XVII) in the presence of hydrogen peroxide and iron salts, according to a procedure described in the literature (Minisci et al. Tetrahedron, 1985, 41, 4157). This method works best when R and R2 together constitute a fused aromatic heterocycle or fused aromatic carbocycle. In those cases that Z is CH and R and R do not form a fused aromatic, an isomeric side product in which Z is CCONHR can be formed and, if so formed, is removed from the desired product by chromatography. Method D

O
XVil
,RD
H
FeS04l H202

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General Method E - The compounds of formula I-E-l and I-E-2 in which R1, R2, R6, M, X, Y, G3, and Z are defined as above, q is 0 or 1, and R3 is lower alkyl are conveniently prepared via a reaction sequence as shown in Method E. Thus, pyridine substituted pyridazines 'or pyridines (I-D-l) are functionalized into substituted 2-alkoxycarbonyl pyridines of formula (I-E-l) by the use of monoalkyloxalates (XVIII) in the presence of S2CV2, acid and catalytic amounts of AgNC>3, according to a procedure described in the literature (Coppa, F. et al., Tetrahedron Letters, 1992, 33 (21), 3057). Compounds of formula I-E-l in which R3 is H are then formed by hydrolysis of the ester with a base such as sodium hydroxide in methanol / water. Compounds of formula I-E-2 in which the Rs groups are independently defined as above, but especially including those compounds in which neither R6 is H, are conveniently prepared from the acid (I-E-l, R3 = H) by treatment with amine XIX in the presence of a coupling agent such as DCC (dicyclohexylcarbodiimide). This method works best when R and R together constitute a fused aromatic heterocycle or fused aromatic carbocycle. In those cases that Z is CH and R' and R2 do not form a fused aromatic, an isomeric side product in which Z is CCO2R3 can be formed in the first step and, if so formed, is removed from the desired product by chromatography.
Method E

General Method F - The compounds of formula I-F in which M, Q2 and X are defined as above, m is an integer of 1 - 5, and R1 and R2 together with the carbons to which they are attached form a fused 5-membered ring aromatic heterocycle can be prepared via a reaction sequence as shown in method F. The readily available heterocyclylcarboxylic acid starting material XX is reacted with butyl lithium followed by dimethylforrnamide to yield the aldehyde with structure XXI. Reaction of XXI with
38

WO 01/23375

PCT/US0O/26500

hydrazine yields pyridazlnone XXII. , Treatment of XXII with a suitable halogenating agent such as POCl3, POBr3, PCI5, PBr5 or thionyl chloride yields a halo intermediate which is reacted with nucleophile of formula V in refluxing alcohol to furnish intermediate compound of formula XXIII. Such condensations can also be done in a melt free of solvent and can be catalyzed by acids such as HC1 or bases such as triefhylamine or 1,8-diazobicyclo[5.4.0]undec-7-ene (DBU). Alternatively, reagent V can be condensed with intermediate XXII be heating the two components with P2O5 in a melt to yield XXII. This last method is especially effective when X is an amine linker. Formation and alkylation of the Reissert compound XXIII with halide XXIV is done as described by the general method of F.D. Popp, Heterocycles, 1980, 14, 1033 to yield the intermediate of structure XXV. Treatment of XXV with base then yields invention compound I-F. Method F

1)BuLi R 2) DMF R2
XXI

2n4
N,H.

XXII

XXIII

XXIII

hal
(CH2)n Q2
Base
XXIV
XXV
General Method G - The compounds of formula I-G in which M, Q2 and X are defined as above, m is an integer of 1 — 4, and R1 and R together with the carbons to which they are attached form a fused 5-membered ring aromatic heterocycle can be prepared via a reaction sequence as shown in method G. Aldehyde XXI, from method F, can be reduced with sodium borohydride to yield a hyroxyacid which is lactonized using methods well known to those skilled in the art such as with toluenesulfonyl chloride to yield lactone XXVI. Condensation of intermediate XXVI with aldehyde XIII in the
39

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presence of a base such as sodium methoxide usually in a solvent such as methanol under reflux yields an intermediate of structure XXVII. Reaction of XXVII with hydrazine or preferably with hydrazine hydrate at a temperature of 100 - 150°C leads to an intermediate of structure XXVIII. Conversion of intermediate XXVIII to invention compound of structure I-G is done by methods as described in method C by using XXVIII rather than XV. '

XXI

1)NaBH4 2) TsCI

XXVI

O
(CH2)5H
Q2 XIII Base

R H XXVII
(CHs^H Q2

N,H,

General Method H - The compounds of formula I-H in which the R1, R , M, X,
R6, q and G3 are defined as above are conveniently prepared via a reaction sequence as shown in Method H. Thus the methods described in Martin, I; Anvelt, J.; Vares, L.; Kuehn, I.; Claesson, A. Acta Chem. Scand. 1995, 49, 230-232 or those of methods D or E above by substituting readily available pyridine-4-carboxylic ester XXX for I-D-l are used to convert XXX into XXXI. Reduction of the ester as described-by Martin, et al. above is next done with a mild reducing agent such as NaBHU such that the amide substiruent is left unchanged to yield alcohol XXXII. This alcohol is then heated with a base such as DBU or CsC04 with halopyridazine VI from method A under anhydrous conditions to yield the invention compound with formula I-H.
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Method H

VI hal -
General Method I - Invention compounds having formula I-I in which the R1, R2, M, X, R6, q, and G3 are defined as above and W is a bond or -CH2- are conveniently prepared via a reaction sequence as shown in Method I. This method is especially useful when q is 1 and XXXIII is 4-chloropyridine. Alternatively, other 4-halopyridines such as 4-fluoropyridine or 4-bromopyndine can be used in this process. Thus readily available 4-halopyridines XXXIII are converted to intermediates of formula XXXIV by using the general procedures of methods D or E above by substituting the 4-halopyridine for I-D-l. Reaction of XXXIV with either potassium or sodium hydrogen sulfide yields a thiol having formula XXXV. Alternatively, the alcohol function of intermediate XXXII from method H is converted to a leaving group by reaction with methanesulfonyl chloride and a suitable base such as triethylamine in the cold such that polymeric material formation is minimized and the resultant intermediate is reacted with either potassium or sodium hydrogen sulfide to. yield a thiol having formula XXXVI. Either thiol have formula XXXV or formula XXXVI is reacted with intermediate VI from method A and a suitable base such as diisopropylethylamine or CSCO4 in DMF or other suitable anhydrous solvent or in the absence of solvent to yield I-D-9.
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Method I

General Method J - Invention compounds such as those having formula I-J-l or I-J-2 in which the R1, R2, M, X, W, and G3 are defined as above and having a sulfoxide or
5 sulfone within the structure are conveniently prepared via a reaction sequence as shown in Method J. Reaction of compounds of this invention that contain a thioether group either as part, of a substituent G1, G3, or G4or as part of Y as shown in the representative structure I-I from Method I can be converted to the invention compounds with a sulfoxide moiety such as I-J-l by treatment with one equivalent of m-chloroperbenzoic acid in methylene 10 chloride or chloroform (MCPBA, Synth. Commun., 26, 10, 1913-1920, 1996) or by treatment with sodium periodate in methanol/water at between 0 °C and room temperature (J. Org. Chem., 58, 25, 6996-7000, 1993). The expected side products consisting of mixtures of various N oxides and the sulfone I-J-2 can be removed by chromatography. The sulfone I-J-2 is obtained by the use of an additional equivalent of MCPBA or 15 preferably by use of potassium permanganate in acetic acid/water (Eur. J. Med. Chem. Then, 21, -1, 5-8, 1986) or by use of hydrogen peroxide in acetic acid (Chem. Heterocycl. Compd., 15, 1085-1088, 1979). In those cases that unwanted N oxides become a significant product, they can be converted back to the desired sulfoxides or sulfones by
42

General Method K - Invention compounds having formula I-K in which the R1, R2, M, X, and Q1 are defined as above are conveniently prepared via a reaction sequence • as shown in Method K. One skilled in the art prepares starting materials of structure XXXVII by methods known in the literature. For example XXXVII wherein R.1 and R2 10 together with the carbons to which they are attached form a 2,3-substituted thiophene, furan, pyrrole, cyclopentadienyl, oxazole or thiazole are prepared using the general chemistry given in J. Org. Chem., 1981, 46, 211 and hydrolizing the initially formed tert-butyl ester with trifluoroacetic acid. The pyrazole starting material can be prepared by reacting 2-oxo-3-pentyn-l,5-dioic acid (J. Chem. Phys. 1974, 60, 1597) with 15 diazomethane. The starting material wherein R and R together with the carbons to which they are attached form a phenyl are prepared by the methods of Cymerman-Craig et al., Aust. J. Chem. 1956, 9, 222, 225. Compounds of formula XXXVII in which R' and R2 are lower alkyl are conveniently prepared according to procedures given in patent CH 482415 (Chem. Abstr. 12026lu, 1970). The crude diacid of formula XXXVII is 20 subsequently treated with hydrazine to furnish pyridazinone XXXVIII (for specific reaction conditions see Vaughn, W. R.; Baird, S. L. J. Am. Chem. Soc. 1946 68 1314). Pyridazinone XXXVIII is. treated with a chlorinating agent such as phosphorous oxychloride to yield an intermediate dichloro species which undergoes hydrolysis upon aqueous workup to furnish chloropyridazine XXXIX. Chloro acid XXXIX is treated with 25 . a nucleophile of formula V in the presence of a base such "as sodium hydride in a solvent such as DMF or in the absence of a solvent. The resultant acid XXXX is reduced with a reducing agent such as BFfj'THF according to the procedure of Till ey, J. W.; Coffen D. L. Schaer, B. H.; Lind,' J. J. Org. Chem. 1987 52 2469. Product alcohol XXXXI is reacted
43

x'M
i
hal Q1
XXXXII
BH,*THF
k.
ha! = F C.\ Rr ,
-K
OH XXXXI
General Method L - Invention compounds having formula I-L in which the R , R2, M, X, and Q1 are defined as above are conveniently prepared via a reaction sequence as shown in Method L. Thus alcohol of formula XXXXI from method K is reacted with methanesulfonyl chloride in the presence of a suitable base followed by potassium or sodium hydrogen sulfide to yield thiol XXXXIII. The thiol is then reacted with 4-halopyridine XXXXII from method K in the presence of a suitable base such as triethylamine to yield invention compound I-K. Alternatively, XXXXI is converted to halo intermediate of formula XXXXIV by methods well known to those skilled in the art and the halide is reacted with thiol XXXXV to yield I-K. Intermediate XXXXIV can also be converted to intermediate XXXXIII by treatment with KHS or.NaHS. Reagents
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XXXXV are either commercially available such as 4-mercaptopyridine or can be prepared by one skilled in the art such as by method I above.

Step 1: Preparation of Intermediate A: A mixture of 2.90 g, 19.07mMol of isocarbostyiil and 14.40 g, 33.68mMol of phosphorus pentabromide were allowed to melt together at 140 °C. The melt turned into a red liquid and after about 10 minutes the reaction mixture solidified and was cooled. The reaction mixture was crushed up and dumped into ice water. The resulting solid was filtered and air-dried, wt. 5.50 g, 96% yield, mp.=94-96°. Rf=0.66 in 40% ethyl acetate in hexanes.
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Br Step 2: A mixture of 1.00 g, 3.49 mMol of 1,4-dibromoisoquinoline (Intermediate A) from step 1 and 4-chloroaniline were melted together at 140°. The reaction mixture turned into a deep red liquid and after about 10 minutes the reaction mixture solidified and was done. The reaction mixture was broken up and triturated with a 50/50 methanol/THF mixture then filtered and air dried without further purification, wt. 0.75 g, 64.4%, mp.=260-263°. Rf=0.58 in 40% ethyl acetate in hexanes.
XI

Step 3: A mixture of 0.05 g, 0.1498 mMol of l-(4-chloroaniline)-4-
10 bromoisoquiniline and 0.02 g, 0.18mMol of 4-mercaptopyridine were combined and
melted together at 140° for about 10 minutes. The resulting reaction mixture was purified
on a 1000 micron prep plate using 5% methanol in hexanes as the solvent, wt. 0.0103 g,
19% yield, mp. 192-195°. R(=0.50 in 40% ethyl acetate in hexanes.
15
46
Example 2: Preparation of l-(indan-5-ylamino)-4-(4-pyridylthio)isoqumoline

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The procedure used for the preparation of Example 1 was used to prepare the title compound by substituting 5-aminoindane for 4-chloroaniline in step 2. Mp. 100-103°, TLC RpO.40 (40% ethyl acetate in hexanes).
^ Example 3: Preparation of l-( benzofhiazoI-6-ylamino)-4-(4-
pyridylthio)isoquinoline

-N
The procedure used for the preparation of Example 1 was used to prepare the title
compound by substituting 6-aminobenzothiazole for 4-chloroaniline in step 2.
10 TLC Rf 0.36 (5%methanol/methylene chloride); MS=387
Example 4: Preparation of l-(4-chIorophenylamino)-4-(4-
pyridyImcthyI)isoquinolme

Step 3: A mixture of 4-chloroaniline (178 mg, 1.40 mmol), phosphorus pentoxide (396 mg, 1.40 mmol) and triethylamine hydrochloride (193 mg, 1.40 mmol) was heated and stirred under argon at 200 C for 1.5 h or until a homogenous melt has formed. To the melt was added starting material (82 mg, 0.35 mmol). The reaction mixture was stirred at 200 0C for 2 h. The resultant solid black mass was cooled to 100 °C. Methanol (5 mL) and water (10 mL) were added and the reaction mixture was sonicated until the black mass had become soluble. Dichloromethane (40 mL) was added and concentrated ammonia (~2 mL) was added to adjust the mixture to pH = 10. The organic layer was separated, and the aqueous layer was extracted with dichloromethane (3 x 20 mL). The combined organic layers were dried over MgS04, filtered, and concentrated. Purification
48

Step 1A: Alternatively, - 3-thiophenecarboxylic acid rather than 2-thiophenecarboxylic acid has been used in step 1 to yield the same product.
a
C02Me
C02Me
Step 2: A 1L, round-bottomed flask was equipped with a stir bar and reflux condenser. To the flask was added the product of step 1 (62 g, 360 mmol) in MeOH (500 mL) with a catalytic amount of H2SO4 (~5 mL). The reaction was heated to reflux, and stirred for 24 h. The reaction was cooled to rt and concentrated rotary evaporation. The brown mixture was purified by silica gel chromatography (Hexane-EtOAc 80:20 gradient to 60:40). Desired dimethyl thiophene-2,3-dicarboxylate was obtained (21.2 g, 106 mmol; 31% yield); 'ii NMR
O
O Step 3: A 250mL, round-bottomed flask was equipped with a stir bar and reflux condenser. To the flask was added the product of step 2 (16 g, 80 mmol), hydrazine hydrate (6.6 mL, 213 mmol), and EtOH (77 mL) and refluxed for 2.5 h. The reaction was cooled to rt and concentrated by rotary evaporation. Water (50 mL) was added and the filtrate was separated from the insoluble solids. The aqueous layer was concentrated by rotary evaporation to give a pale yellow solid. The solid was dried in a vacuum oven
overnight at 50 C. Desired thieno[2,3-d]pyridazin-4,7-dione was obtained (12 g, 71 mmol; 89% yield); "ii NMR (DMSO-d6) 7.85 (d, /= 5.1, 1), 7.42 (d, J= 5.1, 1); ES MS (M+H)+= 169; TLC (dichloromethane-MeOH, 60:40); Rf = 0.58.

Step 4: Preparation of Intermediate B: A 250 mL, round-bottomed flask was equipped with a stir bar and reflux condenser. To the flask was added the product of step 3 (2.5 g, 14.8 mmol), phosphorus oxychloride (45 mL, 481 mmol), and pyridine (4.57 mL, 55 mmol) and refluxed for 2.5 h. The reaction was cooled to rt and poured over ice. The mixture was separated and the aqueous layer was extracted with chloroform (4 x 75 mL).
51

15 Step 6: A 150 mL, round-bottomed flask was equipped with a stir bar and reflux
condenser. To the flask was added the product of step 5 (0.33 g, 1.1 mmol), 4-pyridylcarbinol (1.2 g, 11.2 mmol) in DBU (2.5 mL, 16.7 mmol) and the mixture was heated to 125 °C for 24 hours. EtOAc (10 mL) was added to the reaction while hot and then the reaction was poured into water (10 mL). The layers were separated and the
20 aqueous layer was extracted with EtOAc (3x10 mL). The organic layers were combined, dried (MgS04) and concentrated by rotary evaporation. The resulting mixture was purified by silica gel chromatography (dichloromethane-rnethanol-acetone, 90:5:5) to give a pale yellow solid. The desired title compound was obtained (0.03.g, 0.08 mmol; 7.3% yield);
52

O
NH NH
O Step 3: A 500 mL round bottomed flask fitted with an argon inlet, a reflux condenser, and a stir bar was charged with dimethyl furan-2,3-dicarboxylate (44 g, 236 mmol) dissolved in EtOH (250 mL). Hydrazine hydrate (55 % N2H4, 40 mL, 3.0 mmol) 5 was added to the solution, and the reaction mixture was warmed to reflux. A yellow solid slowly precipitated over the course of 5.5 h, at which point the mixture was cooled to room temperature. The volatiles were removed under reduced pressure to furnish a yellow paste which was suspended in water and filtered. The yellow solid was washed with water and transferred to a 500 mL round bottomed flask fitted with an argon inlet, a reflux 10 condenser, and a stir bar. The solid was suspended in aqueous HCl (2N, 200 mL), and the mixture was warmed to reflux. After heating for 4 h, the orange slurry was cooled to room temperature and filtered. The solid was washed thoroughly with water and dried under vacuum to yield 4,7-dioxo[2,3-Gf]furopyridazine as an orange solid (21.5 g, 60 %). 'HNMR(300MHz, rfs-DMSO) 5 7.00 (d,'.7=2.1, 1), 8.19 (d,/- 2.1, 1H), 11.7 (bs, 2H).
CI

A B
Step 5: A 100 mL round bottomed flask fitted with a stir bar, an argon inlet, and a reflux condenser was charged with the product of Step 4 (1.50 g, 7.98 mmol) dissolved in ethanol (40 mL). Chloroaniline was added to this mixture (1.02 g, 7.98 mmol), and the resultant suspension was warmed to reflux. After heating for 4 h, the mixture was concentrated by rotary evaporation. The crude orange solid was applied to the top of a flash column and eluted with CH2Cl2/MeOH 97:3 to afford a mixture of 4-chloro-7-[JV-(4-chlorophenyl)amino][2,3-d]furopyridazine and 7-chloro-4-[7V-(4-chlorophenyl)amino]-[2,3-
Xi

15
20

Step 6: A 25 mL round bottomed flask was fitted with an argon inlet, a stir bar, and a reflux condenser. The product of step 5 (400 mg, 1.4 mmol) was combined with 4-pyridylcarbinol (782 mg, 7.17 mmol) and l,8-diazabicyclo[5.4.0]undec-7-ene (2.5 mL 16.7 mmol), and the slurry was wanned to 125 °C. After stirring for 24, the reaction was cooled, applied directly to the top of a flash column, and eluted with CH2Cl2/MeOH 95:5. The resultant yellow oil was rechromatographed under the same conditions to yield the title, compound as part of a mixture of three components. HPLC separation (Cjg column CH3CN/H20 10:90 gradient to 100:0) furnished the title compound as an off white solid (13.7 mg, 3- %). TLC (CH2Cl2/MeOH 95:5) = 0.19; MP 198 °C; [H NMR (300 MHz,

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CDCI3) 5 5.60 (s, 2), 6.6 (d, J =2.1, 1), 7.18 - 7.20 (m, 2), 7.35 - 7.43 (m, 6), 7.66 (d, J = 2.1, 1)8.54 (d, 7=5.6, 2).
Steps 5A and 6A: Alternatively 4,7-dibromo[2,3-cz]furopyridazine (Intermediate G below) is used to prepared the title compound by following step 5 but substituting the dibromo intermediate for the dichloro intermediate. Step 6A is conducted by melting the two components together in the presence of CsC04 rather than 1,8-diazabicyclo[5.4.0]undec-7-ene. The crude product is purified as above.
Intermediates D to G: Preparation of other bicyclic 4,5-fused-3,6-dihalopyridazines

The general procedures of example 9, steps 2 to 4 are used by substituting the appropriate heterocycledicarboxylic acid for furan-2,3-dicarboxylic acid to yield the substituted dichloropyridazines D to G found in the below table. The dibromofuropyridazine G was prepared using steps 2-3 from example 9 and then conducting step 4' as follows: to 0.50g (3.287 mmol) of the product of step 3 was added 2.83g (6.57 mmol) of phosphorus pentabromide. This was heated to 125 °C. At about 115 °C the reaction mixture melted and then re-solidified before it reached 125 °C. The reaction mixture was cooled and the solid residue was crashed up and dumped into ice water. The resulting solid was then filtered and vacuum dried. wt.=0.75g (82% yield). In several cases the dichloropyridazines are known materials, as indicated by the given reference. All of these dihaloheterocycles can be used to prepare the claimed invention compounds. TABLE

Intermediates I to N: General Method for Preparation of [2-(N-Substituted)aminocarbonyI-4-pyridyI] methanol Intermediates
HO
H(X
NR'IT
OEt

HNR1R2
benzene, reflux

15

20

To a 0 °C solution of the amine 2 (3 equiv) in benzene is added trimefhyl aluminum (3 equiv). Gas evolution is observed and the reaction is then allowed to warm to rt and stir for 1 h. (Lipton, M.F. et al. Org. Synth. Coll. Vol. 6, 1988, 492 or Levin, J.I. et al. Synth. Comm., 1982, 12, 989). The known carbinol 1 (1 equiv, Hadri, A. E.; Leclerc, G. Heterocyclic Chem, 1993, 30, 631) is added to the aluminum reagent and the mixture is heated to reflux for lh. The reaction is quenched with water and concentrated. The crude product is usually purified by silica gel column chromatography (20/1 EtOAc/MeOH) to afford title compound 3. The final products are generally confirmed by LC/MS and NMR spectroscopy.

A 25 mL, 3-necked, round-bottomed flask was equipped with a stir bar and thermometer. To the flask was added the product of Example 8 (0.475 g, 1.29 mmol), iron sulfate heptahydrate (0.179 g, 0.64 mmol), formamide (11.15 mL, 281 mmol) and cone. H2SO4 (0.14 mL). The mixture was stirred for 30 min at rt at which time H202 (0.2 mL, 10 6.44 mmol) was added drop wise to the mixture. The reaction stirred at room temperature for an additional hour and then heated to 55 °C over 30 min. The reaction was kept at this temperature for 3 h and then cooled to rt. An aqueous solution of sodium citrate (0.27M, 1 mL) was added to the reaction and subsequently the layers were separated and the aqueous layer was extracted with EtOAc (4 x 5mL). The organic layers were combined, dried 15 (MgSO^ and concentrated by rotary evaporation. The resulting solid was taken up in hot acetone and separated from any remaining solids by filtration. The filtrate was then concentrated by rotary evaporation and the resulting residue was taken up in hot MeOH and the white solid was collected by filtration. Desired compound (.014 g, 0.034 mmol;
59

XI
-NHCH3 ^K|>~^/NHCH3
O O
. To a suspension of the final product from Example 9 (19.20 g, 54.4 mmol) in N-methylformarnide (200 mL) and distilled water (20 mL) at room temperature was added concentrated H2SO4 (2.9 mL, 54.4 mmol) dropwise. The mixture was stirred until it became a clear solution. To this solution was added FeSO^hbO (1.51 g, 5.43 mmol) in one portion, followed by the addition of hydroxylamine-O-sulfonic acid (HOSA, 1.84 g, 16.3 mmol). The additions of FeS04-7H20 and HOSA were repeated in 10 min. intervals

o
Intermediate H
To a mixture of the Intermediate from Example 9, step 5 (10.0 g, 35.7 mmol), 25 Intermediate H (12.4 g, 74.6 mmol), and 18-crown-6 (0.42 g, 1.59 mmol) in toluene (100 mL) was added KOH powder (4.4 g, 85%, 66.7-mmol) in one portion at room temperature. The reaction mixture was then heated to 85 ± 2 °C under vigorous stirring. The reaction mixture was stirred vigorously at this temperature overnight. After it was cooled to room temperature, toluene solution was decanted off and water (100 mL) was
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added to the gummy residue. The resulting mixture was stirred vigorously until it became a free flowing suspension. The solids were collected by filtration, washed with water (2 x 10 mL), and dried under vacuum at 45 °C for 16 hours. The yellow/brown solids were suspended in acetonitrile (70 mL) and the suspension was stirred at reflux for 2 hours. After it was cooled to room temperature, the solids were collected by filtration, washed with small amount of acetonitrile, and dried under vacuum at 45 °C overnight. The title product was isolated in 46% yield (6.73 g) as a light yellow solid.
10 Example 16: Preparation of 4-(4-chlorophenylamino)-7-(2-aminocarbonyl-4-pyridyImethoxy)furo-[2,3-d]pyridazine

The procedure used for the preparation of Example 14 was used to prepare the title 15 compound by substituting Formamide for N-methylformamide. The reaction was conducted with 500 mg of final product from Example 9 and proportional amounts of solvents and reagents. The crude product was purified by HPLC on a 75x30 mm C18 column and a linear gradient elution from 10 to 100% acetonitrile in water with 0.1% trifluoroacetic acid at 10 ml/min. over 10 min. to yield 18 mg of the title compound as a 20 yellow solid: HPLC (50x4.6 mm YMC CombiScreen® CIS column, linear gradient 10 to 100%o acetonitrile in water with 0.1% trifluoroacetic acid at 3 ml/min. over 5 min., UV detection at 254'nm) 2.35 min. peak; MS ES 396 (M+H)+.
Example 17: Preparation of 4~(4-chlorophenylamino)~7~(benzothiazol-6-25 ylaminp)thieno[2,3-d]pyridazine
63

xx>
20 The procedure used for the preparation of Example 17 was used to prepare the title
compound by substituting 5-aminomdane for 4-chloroaniline. The crude product was purified by flash chromatography on silica gel using 30% ethyl acetate/hexane.as the eluent. The structure of the pure title compound was confirmed by LC/MS and NMR:
64

N
4,7-Dichloro[2,3-J]furopyridazine from step 4 of Example 9 (95 mg, 0.50 mmol) and 5-bromoindoline (100 mg, 0.50 mmol) weie refluxed in 60 mL of absolute ethanol at 95 °C for 2 hrs. The reaction mixture was allowed to cool to room temperature and the precipitate that formed was filtered and washed with isopropyl alcohol, 4.0 N KOH, H20, and hexane, and then dried. The intermediate of about 95% purity (rt = 4.72, (M+H)+ 350) and was used in the next step without further purification. 4-Pyridylcarbinol (28 mg, 0.26 mmol) and sodium hydride (60%, 50 mg, 1.25 mmol) were stirred in 20 mL of anhydrous tetrahydrofuran at 0 °C under Argon for 20 min. and then 44 mg of the above intermediate (0.13 mmol) was added. The reaction was stirred at 0 °C for 2 hrs and the temperature was allowed to rise to room temperature. The mixture was stirred for . another 12 hrs and the solvent was evaporated under reduced pressure. The solid that was obtained was dissolved in 50 mL of dichloromethane and washed with K2CO3 solution and BbO. The organic layer was separated, dried (MgSCU), and evaporated under reduced pressure. The crude product was purified by preparative TLC (Rf = 0.3) on silica gel using dichloromethane/methanol (95:5) as the eluent. The structure of the pure title compound was confirmed by LC/MS and NMR: 'H NMR (CDC13) 8 3.20 (m, 2H), 4.30 ~ 4.50 (m, 2H), 5.60 (s, 2H), 6.9 ~ 8.0 (m, 7 H), 8.60 (m, 2H); LC/MS (M+H)+ 423 rt = 4.49 min.
65

■Ov.
O To a suspension of 4,7~Dichloro[2,3-^furopyridazine from step 4 of Example 9 (400 mg, 2.12 mmol, 1 equiv) and p-anisidine 0-MeOC6H4NH2) (260 mg, 2.12 mmol, 1 equiv) in DME (5 mL) was added water (1 mL). The resulting solution was heated at 50 °C for 48 h. After cooling to rt, the brown precipitate was removed by filtration and the filtrate was concentrated in vacuo to afford the crude product as a brown solid. Trituration of the brown solid with CLI2CI2 furnished 292 mg (50%) of the intermediate 4-(4-methoxyphenylamino)-7chlorofuro-[2,3-d]pyridazine which was confirmed by LC/MS and NMR. A suspension of this intermediate (292 mg, 1.06 mmol, 1 equiv), (2-methylaminocarbonyl-4-pyridyl)methanol (Intermediate H,_529.mg, 3.18 mmol, 3 equiv) and 18-crown-6 (42 mg, 0.16 mmol, 15 mol%) in toluene (4 mL) was stirred at rt for 20 ■min. KOH (178 mg, 3.18 mmol, 3 equiv) was then added and the reaction mixture was heated to 80 °C for 36 h. After cooling to rt, water (10 mL) was added and the mixture was stirred vigorously until a fine white suspension was formed. The suspension was filtered and washed with water and diethyl ether to provide 125 mg (29%) of the desired product as a light yellow solid: (M+H)+ 406; R/= 0.50 (100% EtOAc).
66

The procedure used for the preparation of Example 20 was used to prepare the title 5 compound by substituting 4-pyridylmethanol for (2-meth-ylaminocarbonyl-4-pyridyl)methanol. The pure product was isolated by chromatography on a flash column: (M+H)+ 349; R/= 0.3 (95:5 CH2C12/CH30H).
Example 22: Preparation of 4-(4-methoxyphenylarnmo)-7-(2-aminocarbonyl-4-10 pyridyImcthoxy)furo-[2,3-d]pyridazine

o
The procedure used for the preparation of Example 16 was used to prepare the title
compound by substituting the product of Example 21 for the product from Example 9.
The reaction was conducted with 250 mg of the starting material and proportional amounts
15 of solvents and reagents. The crude product was purified by HPLC on a 75x30 mm CI 8
column and a linear gradient elution from 10 to 100% acetonitrile in water with 0.1%
• trifluoroacetic acid at 10 ml/min. over 10 min. to yield 16 mg of the title compound as a
yellow solid: HPLC (50x4.6 mm YMC CombiScreen® CI8 column, linear gradient 10 to
100% acetonitrile in water with 0.1% trifluoroacetic acid at 3 ml/min. over 5 min., UV
20 detection at 254 nm) 1.98 min. peak; MS ES 392 (M+H)+.
67

Method A-l: Equal equivalents of dichloride (1) and M-NH2 are refluxed in the appropriate amount of absolute ethanol at 95 °C for 2 hrs. The reaction mixture is allowed to cool to room temperature and the precipitate (2) that forms is filtered and washed sequentially with isopropyl alcohol, 4.0 N KOH, H2O, and hexane, and then dried. The filtrate (2) is then reacted with 1.2 equivalent of Q-NH2 in an appropriate amount of n-butyl alcohol at 150 °C for 10 hrs. The reaction is cooled to room temperature before the solvent is evaporated under reduced pressure. The residue is treated with aqueous 4.0 N KOH solution and extracted with dichloromefhane. The organic layer is dried (MgS04) and evaporated. The crude product (3) is purified by preparative thin layer chromatography (TLC) or flash chromatography on silica gel using dichloromethane/methanol (95:5) as the eluent. Final product is confirmed by LC/MS and/or NMR. The invention compounds of Examples 23 - 25, 48, and 76-80 as shown in the below table were prepared by method A-l.

M-NH2 2.2 equiv.
n-BuOH; 150 °C, 10 hrs.

-J»-

NT
NH
M
HNL

20
25

Method A-2: One equivalent of dichloride (1) and 2.2 equivalent of M-NH2 are refluxed in an appropriate amount of n-butanol at 150 °C for 10 hrs. The reaction mixture is allowed to cool to room temperature and the precipitate (4) that forms is filtered and washed sequentially with isopropyl alcohol, 4.0 TV KOH, H20, and hexane, and then dried. The crude product (4) is purified by preparative TLC or flash chromatography on silica gel using dichloromethane/methanol (95:5) as the eluent. Final product is confirmed by LC/MS and/or NMR. The invention compounds of Examples 26 — 33 and 75 as shown in the below table were prepared by method A-2.

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M-NH,
DME/H20, 55 °C, 48h

Q-OH (3)
tol, 18-crown-6, KOH, 80 °C, 36 h

10
15

Method A-3: One equivalent of dichloride (1) and one equivalent of M-NH2 are
suspended in DME (0.3M) and water is added until a solution was formed. The reaction
mixture is heated to 65 °C for 48 h. After cooling to it, the resulting precipitate is filtered
and washed with DME to provide the intermediate product (2) which is confirmed by
LC/MS and NMR.. In some instances, intermediate (2) is further purified by preparative
TLC or washed with other solvents. A suspension of (2) (1 equiv), carbinol (3) (3 equiv),
and 18-crown-6 (10 mol %) in toluene (0.3M) is stirred at it for 10 min. KOH (3 equiv) is
then added and the reaction mixture is heated to 80 °C for 24 h. After cooling to rt, water
is added and the mixture is stirred vigorously until a suspension is formed. The
suspension is filtered and washed with water to provide the desired product (4).
Preparative TLC and/or washing with other solvents is used to further purify final
products in some examples. The final products are assigned by LC/MS and NMR
spectroscopy. Final product is confirmed by LC/MS and/or NMR. The invention
compounds of Examples 34 - 47, 49-74, and 81 — 82D as shown in the below table were
prepared by method A-3.

, the compounds were characterized' by TLC on silica gel plates and the Ry values and solvents are shown. HPLC retention times are given for other examples in this table; a HPLC - electrospray mass spectra (HPLC ES-MS) were obtained using a Hewlett-Packard 1100 HPLC equipped with a quaternary pump, a variable wavelength detector, a YMC 5 Pro CI8 2.0 mm x 23 mm column, and a Finnigan LCQ ion trap mass spectrometer with electrospray ionization. Gradient elution from 90% A to 95% B over 4 minutes was used on the HPLC. Buffer A was. 98% water, 2% Acetonitrile and 0.02% TFA. Buffer B was 98% Acetonitrile, 2% water and 0.018% TFA. Spectra were scanned from 140-1200 amu using a variable ion time according to the number of ions in the source; An HPLC assay 10 with UV peak detection was ran in addition to the HPLC ES-MS experiment and the conditions are: 50x4.6 mm YMC CombiScreen® C18 column, linear gradient 10 to 100%o acetonitrile in water with 0.1% trifluoroacetic acid at 3 ml/min. over 5 min., UV detection at 254 ran; c The product was purified by RP-HPLC on a CI8 column using a water/acetonitrile gradient with added trifluoroacetic acid such that the trifluoroacetate salt 15 was isolated by evaporation of the pure product; d4-pyridylmethanol, as indicated, was ' used in step 2 of method A-l rather than an amine; cFor preparation of 5-amino-2,3-dihydrobenzofurane see Mitchell, H.; Leblanc, Y. /. Org. Chem. 1994, 59, 682-687. f The reference to make the known TBS protected alcohol intermediate is : Parsons, A. F.; Pettifer, R. M. J.Chem. Soc. Perkin Trans. 1, 1998, 651.
>~o o
20 The deprotection of ^^ was accomplished in the following
manner:
Three equiv of a 1.0 Molar solution of TBAF in THF was added to a solution of
the protected alcohol in TFIF (0.05Molar) at rt. The reaction mixture was allowed to stir
at rt for 1 h and was quenched with water followed by extraction with EtOAc. 25
76
SUBSTITUTE SHEET (RULE 26)
Examples 83 - 92: Preparation of Isoquinolincs by Method B-l

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Method B-l: Dibromoisoquinoline (5, 29 mg, 0.1 mmol) Example 1, step 1, and M-NH2 (0.2 mmol) in 8-mL vial were heated in 1 mL of n-butanol at 90 °C for 36 hrs. The mixture was cooled to room temperature and the solvent was evaporated under reduced pressure. 4-Mercaptopyridine (23 mg, 0.2 mmol) and cesium carbonate (67 mg, 5 0.2 mmol) were added to the vial. The mixture was heated at 180 °C for 1 hr and was allowed to cool to room temperature. Methanol (2 mL) was added to the vial and the mixture was sonicated for 10 min and filtered. The methanol solution of reaction mixture was collected and evaporated under reduced pressure. The formation of product was confirmed by LC/MS. The invention compounds of Examples 83 - 92 as shown in the 10 below table were prepared by method B-l.
Compounds that were Prepared by Method B-l

detector, a YMC Pro CI 8 2.0 mm x 23 mm column, and a Finnigan LCQ ion trap mass spectrometer with electrospray ionization. Gradient elutioh from 90% A to 95% B over 4 minutes was used on the HPLC. Buffer A was 98% water, 2% Acetonitrile and 0.02% TFA. Buffer B was 98% Acetonitrile, 2% water and 0.018% TFA. Spectra were scanned from 140-1200 amu using a variable ion time according to the number of ions in the source.
Examples 93 - 105: Preparation of Novel Phthalazine Invention Compounds by Parallel Synthesis
10 Method A-l or A-2, as indicated, were used to prepare the novel phthalimide
invention compounds 93 - 105 from 1,4-dichIorophfhalazine (for preparation see Novartis patent W09.8/35958, 11.02.98) rather than the dichloroheterocyclopyridazines together with the appropriate bicyclic and substituted anilines.

detector, a YMC Pro CI8 2.0 mm x 23 mm column , and a Finnigan LCQ ion trap mass spectrometer with electrospray ionization. Gradient elution from 90% A to 95% B over 4 minutes was used on the HPLC. Buffer A was 98% water, 2% Acetonitrile and 0.02% TFA. Buffer B was 98% Acetonitrile, 2% water and 0.018% TFA. Spectra were scanned from 140-1200 amu using a variable ion time according to the number of ions in the source.
Examples 106 - 114: Preparation of Salts of Example 14.
The product.of Example 14 (1.50 g, 3.66 mmol) was stirred as a slurry in methanol (20 ml) as a solution of toluenesulfonic acid hydrate (0.701 g, 3.67 mmol) in methanol (5 ml plus 5 ml rinse) was added quickly dropwise. All materials dissolved over 5 min to yield a yellow solution. Anhydrous ether (30 ml) was added and stirring was continued for 5 minutes until solid began to precipitate. The resultant mixture was chilled with stirring in an ice/water bath for 45 minutes and then the solid title product (Example 104) was collected by filtration, washed with ether and dried at 55 °C in a vacuum oven until NMR analysis showed a lack of solvents (1.5 hours). Other compounds were prepared in a similar way by using a variety of acids rather than toluenesulfonic acid. Scale up and use of less methanol in the first step generally led to quicker precipitation of salts and a variety of solvents were used rather than ether, as indicated, to help crystalize the individual salts. In some cases the methanol was first removed by evaporation in vacuo. Final drying took between 1.5 hours and several days, depending on the quantity of material and the specific specific acid used.
Salts of Example 14 that were Prepared

was generated from 500 micrograms/ml to 30 micrograms/ml in Lysis Buffer (see above). Standard curve and lysate' samples were added at 5 microliters /well in duplicate to a polystyrene 96-well plate. Using the Sigma Plasma Hemoglobin Kit (Sigma Chemical Co., St. Louis, MO), TMB substrate was reconstituted in 50 mis room temperature acetic 5 acid solution. One hundred microliters of substrate was added to each well, followed by 100 microliters /well of Hydrogen Peroxide Solution at room temperature. The plate was incubated at room temperature for 10 minutes.
Optical densities were determined spectrophotometrically at 600 nm in a 96-well , plate reader, SpectraMax 250 Microplate Spectrophotometer System (Molecular Devices, 10 Sunnyvale, CA). Background Lysis Buffer readings were subtracted from all wells.
Total sample hemoglobin content was calculated according to the following equation:
Total Hemoglobin = (Sample Lysate Volume) x (Hemoglobin Concentration) 15
The average Total Hemoglobin of Matrigel samples without cells was subtracted from each Total Hemoglobin Matrigel sample with cells. Percent inhibition was calculated according to the following equation:
20 % Inhibition = (Average Total Hemoglobin Drug-Treated Tumor Lysates) X 100
(Average Total Hemoglobin Non-Treated Tumor Lysates)
Example 8 showed significant activity in this assay at 100 and 300 mg/kg po sid with > 60% inhibition of total hemoglobin content of the Matrigel samples from the dosed 25 animals vs. those from vehicle control animals. The other examplary materials were not tested in this model.
Other embodiments of the invention will be apparent to the skilled in the art from a consideration of this specification or practice of the invention disclosed herein. 30 It is intended that the specification and examples be considered as exemplary only, with the true scope and spirit of the invention being indicated by the following claims.
87

• -OCON(R6)2 ;
R3is H or (Ci-Cs)alkyl;
R6 is independently selected from the group consisting of
• H; and
• (Ci-Cs)alkyl;
R4 is H, halogen, or (Ci-Csjalkyl;
p is 0, 1, or 2;
X is selected from the group consisting of O, S, and NR3;
Y is selected from the group consisting of
• -O-;
• -S- ; and
• -NH- ;
Z is N or CR4; qis 1;
G3 is a bivalent moiety selected from the group consisting of
• bivalent bridges of structure T2=T2-T3 :
wherein
each T2 independently represents N or CH ; and
T3 represents S, O, or NR3 ; and
the terminal T2 is bound to L, and T3 is bound to D,
forming a 5-membered fused ring;

A and D are CH;
B and E are CH;
LisCH;
with the proviso that the resulting phenyl ring bears as a G3 substituent said bivalent bridge of structure T2=T2-T3;
J is a ring selected from the group consisting of
• phenyl; and
• pyridyl;
q' represents the number of substituents G4 on ring J and is 0, 1, 2, or 3 and
G4 is a monovalent or bivalent moiety selected from the group consisting of
• -N(R6)2 ;
• halogen;
• (Ci-C5)alkyl;
• OR6; and
• fused ring-forming bivalent bridges attached to and connecting
adjacent positions of ring J, said bridges having the structures: a)
Tv
T3 wherein
each T2 independently represents N or CH;
T3 represents S, O, or NR3; wherein
binding to ring J is achieved via terminal atoms T2 and T3;
hi
11

^ 6 TV
Tfa or
/ T6
T5 T5^
wherein
each T5 and T6 independently represents O, S, or CF2 ; and binding to ring J is achieved via terminal atoms T4 or T5 ; with the provisos that:
i) a bridge comprising T5 and T6 atoms may contain a
maximum of two heteroatoms O or S; and ii) in a bridge comprising T5 and T6 atoms, when one T5 group and one T6 group are O atoms, or two T6 groups are O atoms, said O atoms are separated by at least one carbon atom;
when G4 is an alkyl group located on ring J adjacent to the linkage -(CR42JP- , and X is NR3 wherein R3 is an alkyl substituent, then G4 and the alkyl substituent R3 on X may be joined to form a bridge of structure -(CH2)P' - wherein p' is 2 or 3 with the proviso that the sum of p and p' is 2 or 3 resulting in formation of a nitrogen-containing ring of 5 or 6 members;
and with the further provisos that:
- in G1 and G4, when two groups R6 are each alkyl and located on the same N atom they may be linked by a bond, an O, an S, or NR3 to form a N-containing heterocycle of 5 - 7 ring atoms;
or a pharmaceutically acceptable salt or prodrug thereof.
2. A compound as claimed in claim 1 wherein, in the ring comprising A, B, D, E, and L and a bivalent bridge of structure T2=T2-T3 , the terminal T2 represents N and the T3 unit of said bridge represents S, O, or NR3.
°l\y

3. A pharmaceutical composition comprising a compound as claimed in claim 1 and a pharmaceutically acceptable carrier.
4. A method of treating a mammal having a condition characterized by abnormal angiogenesis or hyperpermiability processes, comprising administering to said mammal an amount of a compound of claim 6 which is effective to treat said condition.
5. The method as claimed in claim 4, wherein said condition is tumor growth; retinopathy, including diabetic retinopathy, ischemic retinal-vein occlusion, retinopathy of prematurity, and age-related macular degeneration; rheumatoid arthritis; psoriasis; or a bullous disorder associated with subepidermal blister formation, including bullous pemphigoid, erythema multiforme, and dermatitis herpetiformis.
6. A compound selected from the group consisting of